A fundamental understanding of the relationship between cotton fiber strength (or tenacity)/elongation and structure is important to help cotton breeders modify varieties for enhanced end-use qualities. In this study, the Stelometer instrument was used to measure the bundle fiber tenacity and elongation properties of different cotton fibers. This instrument is the traditional fiber strength reference method and could be still preferred as a screening tool owing to its significant low cost and portability. Fiber crystallinity (CI IR ) and maturity (M IR ) were characterized by the previously proposed attenuated total reflection (ATR)-based Fourier transform infrared protocol that has microsampling capability and is suitable for the tiny Stelometer breakage specimens (2 $ 5 mg), which cannot be readily analyzed by a conventional X-ray diffraction pattern. Relative to the distinctive increase in fiber tenacity with either CI IR or M IR for Pima fibers (Gossypium barbadense), there was an unclear trend between the two for Upland fibers (G. hirsutum). Although fiber elongation increases with elevated CI IR and M IR for Pima fibers, it generally decreases as CI IR and M IR increase for Upland fibers. Furthermore, small sets of Upland fibers with known varieties and growth areas were examined, and their responses to both CI IR and M IR are discussed briefly.One of the essential attributes of cotton fibers is strength, which is required to manufacture quality goods for consumers. Over the years, a number of techniques, such as the Mantis single-fiber tester, 1,2 the Instron tensile tester, 3 the Favimat single-fiber tester, 4,5 Fibrotest, 6 the Stelometer bundle tester, 2,5-9 and the high volume instrument (HVI), 2,5,6,9,10 have been developed to measure cotton strength by testing either a single fiber or a bundle of fibers. Comparisons between two or more independent strength tests have been attempted by different investigators. 2,5,6,9 Thibodeaux et al. 2 determined the relationship between the single-fiber (Mantis) strength and two-bundle fiber (Stelometer and HVI) strength and found that both the Stelometer strength and HVI bundle strength are linearly proportional to the ratio of the average Mantis breaking load to the square of the average fiber ribbon width. By analyzing the results of either single-fiber testing from the Favimat or bulky fiber measurements from the HVI and Stelometer, Delhom et al. 5 reported that single-fiber testing results in higher mean values than bundle testing. In a more recent investigation, Cui et al. 6 observed that the Fibrotest absolute strength is lower than the Stelometer strength. There was also a report on the general conversion between the two bundle strength readings from the Stelometer and HVI measurements. 9